The role of dietary niacin intake and the adenosine-5'-diphosphate-ribosyl cyclase enzyme CD38 in spatial learning ability: is cyclic adenosine diphosphate ribose the link between diet and behaviour?

The pyridine nucleotide NAD+ is derived from dietary niacin and serves as the substrate for the synthesis of cyclic ADP-ribose (cADPR), an intracellular Ca signalling molecule that plays an important role in synaptic plasticity in the hippocampus, a region of the brain involved in spatial learning. cADPR is formed in part via the activity of the ADP-ribosyl cyclase enzyme CD38, which is widespread throughout the brain. In the present review, current evidence of the relationship between dietary niacin and behaviour is presented following investigations of the effect of niacin deficiency, pharmacological nicotinamide supplementation and CD38 gene deletion on brain nucleotides and spatial learning ability in mice and rats. In young male rats, both niacin deficiency and nicotinamide supplementation significantly altered brain NAD+ and cADPR, both of which were inversely correlated with spatial learning ability. These results were consistent across three different models of niacin deficiency (pair feeding, partially restricted feeding and niacin recovery). Similar changes in spatial learning ability were observed in Cd38- / - mice, which also showed decreases in brain cADPR. These findings suggest an inverse relationship between spatial learning ability, dietary niacin intake and cADPR, although a direct link between cADPR and spatial learning ability is still missing. Dietary niacin may therefore play a role in the molecular events regulating learning performance, and further investigations of niacin intake, CD38 and cADPR may help identify potential molecular targets for clinical intervention to enhance learning and prevent or reverse cognitive decline.

Rat models of caloric intake and activity: relationships to animal physiology and human health.

Every rodent experiment is based on important parameters concerning the levels of caloric intake and physical activity. In many cases, these decisions are not made consciously, but are based on traditional models. For experimental models directed at the study of caloric intake and activity, the selection of parameters is usually aimed at modeling human conditions, the ultimate goal of which is to gain insight into the pathophysiology of the disease process in man. In each model, it is important to understand the influence of diet, exercise, and genetic background on physiology and the development of disease states. Along the continuum of energy intake from caloric restriction to high-fat feeding, and of energy output from total inactivity to forced exercise, a number of models are used to study different disease states. In this paper, we will evaluate the influence of the quantity and composition of diet and exercise in several animal models, and will discuss how each model can be applied to various human conditions. This review will be limited to traditional models using the rat as the experimental animal, and although it is not an exhaustive list, the models presented are those most commonly represented in the literature. We will also review the mechanisms by which each affects rat physiology, and will compare these to the analogous mechanisms in the modeled human disease state. We hope that the information presented here will help researchers make choices among the available models and will encourage discussion on the interpretation and extrapolation of results obtained from traditional and novel rodent experiments on diet, exercise, and chronic disease.

The guinea-pig is a poor animal model for studies of niacin deficiency and presents challenges in any study using purified diets.

The guinea-pig was previously reported as being sensitive to a niacin-deficient (ND), high-protein diet, suggesting that it is a suitable model for the low tryptophan to NAD+ conversion observed in human subjects. However, these studies were based on growth rates and mortality. The objective of the present study was to determine whether guinea-pigs are suitable for ND studies based on measurements of blood and bone marrow NAD+. Using a 20 % casein diet, ND decreased blood NAD+ after 4 weeks, but this parameter returned to normal after 9 weeks of feeding, while bone marrow was decreased by 35 % at this time point. Using a 15 % casein diet, 7 weeks of ND caused 44 and 42 % decreases in blood and bone marrow NAD+. Using a 10 % casein diet, ND decreased NAD+ by 32 % in blood and 62 % in bone marrow at 7 weeks. Growth rates were directly related to the dietary tryptophan content, with the lowest growth rates seen with the 10 % casein diet. Changes in guinea-pig NAD+ are comparable with the rat model at similar levels of dietary tryptophan, while mortality rates were dramatically higher in the guinea-pig model. The present study concludes that mortality in ND guinea-pigs is not indicative of poor tryptophan conversion, but is due to environmental stresses in guinea-pigs that are not observed with rats. We conclude that guinea-pigs are not suitable for research on niacin deficiency and they present challenges for any study requiring purified diets and wire-bottomed cages.

Water maze performance in young male Long-Evans rats is inversely affected by dietary intakes of niacin and may be linked to levels of the NAD+ metabolite cADPR.

Niacin is converted in tissues to NAD(+), which is required for synthesis of the intracellular calcium signaling molecule cyclic ADP-ribose (cADPR). cADPR is involved in many aspects of cognitive function, including long-term depression, in the hippocampus, a brain region that regulates spatial learning ability. The objective of this study was to determine whether niacin deficiency and pharmacological nicotinamide supplementation have an effect on spatial learning ability in young male Long-Evans rats as assessed by the Morris Water Maze, and whether brain NAD(+) and cADPR are modified by dietary niacin intake. We investigated 3 models of niacin deficiency: niacin deficient (ND) vs. pair fed (PF), ND vs. partially feed restricted (PFR), and ND vs. niacin recovered (REC). ND rats showed an improvement in spatial learning ability relative to PF, PFR, and REC rats. ND rats also showed a decrease in both NAD(+) and cADPR relative to PF and REC rats. We also investigated 1 model of pharmacological supplementation, niacin-supplemented vs. control. The niacin-supplemented group showed a small but significant spatial learning impairment relative to controls, and an increase in brain cADPR and NAD(+). Changes in neural function related to the NAD(+) associated calcium signaling molecule, cADPR, may be the link between diet and behavior.

Modifications to increase the efficiency of the fluorometric cycling assay for cyclic ADP-ribose.

Cyclic ADP-ribose (cADPR) is an intracellular messenger that triggers the release of calcium ions from intracellular stores in a variety of cell types. The fluorometric cycling assay has become the preferred method for measuring cADPR due to its high level of sensitivity (in the sub-nanomolar range) and its use of commercially available reagents. Additionally, the assay is performed in multiwell plates, making it suitable for high throughput screening using a fluorescence plate reader. The findings reported in this paper present several problems that may be encountered during various stages of the assay, and provide solutions to these problems. Modifications to the assay address reduced recovery of sample and cADPR with removal of perchloric acid (PCA) using organic solvent, reduction in diaphorase activity with heat treatment, and effects on resorufin fluorescence by pH range. Using these modifications, we report an increase of approximately 15% in recovery of brain cADPR, and show that between-subject variability is greatly reduced. We hope that these observations will encourage more widespread application of this valuable assay.

Decreased cADPR and increased NAD+ in the Cd38-/- mouse.

CD38 is a type II glycoprotein that catalyzes the formation of cyclic ADP-ribose (cADPR), an intracellular calcium signalling molecule, from nicotinamide adenine dinucleotide (NAD(+)). Using a modified version of the fluorimetric cycling assay for cADPR which reduces between-subject variability, we report significant decreases in brain and lung cADPR, which although similar to previously published values, showed much less individual variation. The reduced variation within each group suggests that the range of cADPR is narrower than previously thought, and that the regulatory mechanisms controlling these levels are more finely tuned. We also report significant increases in brain, lung, and kidney NAD(+) in the Cd38(-/-) mouse, and provide the first experimental demonstration of the proximate relationship between CD38 and NAD(+).

Use of salient and non-salient visuospatial cues by rats in the Morris Water Maze.

In the Morris Water Maze (MWM), an animal learns the location of a hidden platform relative to distal visual cues in a process known as spatial learning. The visual cues used in MWM experiments are invariably salient in nature, and non-salient cues, such as subtle environmental variations, have not traditionally been considered to play a significant role. However, the role of non-salient cues in spatial navigation has not been adequately investigated experimentally. The objective of this experiment was therefore to determine the relative contribution of salient and non-salient visual cues to spatial navigation in the MWM. Animals were presented with an environment containing both types of visual cues, and were tested in three successive phases of water maze testing, each with a new platform location. Probe tests were used to assess spatial accuracy, and several cue variation trials were run in which both salient and non-salient visual cues were manipulated. It was observed that removal of the salient visual cues did not cause a significant deterioration in performance unless accompanied by disruption of the non-salient visual cues, and that spatial navigation was unimpaired when only the salient visual cues were removed from view. This suggests that during place learning in Long-Evans rats, non-salient visual cues may play a dominant role, at least when salient cue presentation is limited to four cues.

Effect of randomized supplementation with high dose olive, flax or fish oil on serum phospholipid fatty acid levels in adults with attention deficit hyperactivity disorder.

Dietary intake of omega-3 fatty acids has been positively correlated with cardiovascular and neuropsychiatric health in several studies. The high seafood intake by the Japanese and Greenland Inuit has resulted in low ratios of the omega-6 fatty acid arachidonic acid (AA, 20:4n-6) to eicosapentaenoic acid (EPA, 20:5n-3), with the Japanese showing AA:EPA ratios of approximately 1.7 and the Greenland Eskimos showing ratios of approximately 0.14. It was the objective of this study to determine the effect of supplementation with high doses (60 g) of flax and fish oils on the blood phospholipid (PL) fatty acid status, and AA/EPA ratio of individuals with Attention Deficit Hyperactivity Disorder (ADHD), commonly associated with decreased blood omega-3 fatty acid levels. Thirty adults with ADHD were randomized to 12 weeks of supplementation with olive oil (< 1% omega-3 fatty acids), flax oil (source of alpha-linolenic acid; 18:3n-3; alpha-LNA) or fish oil (source of EPA and docosahexaenoic acid; 22:6n-3; DHA). Serum PL fatty acid levels were determined at baseline and at 12 weeks. Flax oil supplementation resulted in an increase in alpha-LNA and a slight decrease in the ratio of AA/EPA, while fish oil supplementation resulted in increases in EPA, DHA and total omega-3 fatty acids and a decrease in the AA/EPA ratio to values seen in the Japanese population. These data suggest that in order to increase levels of EPA and DHA in adults with ADHD, and decrease the AA/EPA ratio to levels seen in high fish consuming populations, high dose fish oil may be preferable to high dose flax oil. Future study is warranted to determine whether correction of low levels of long-chain omega-3 fatty acids is of therapeutic benefit in this population.

Blood phospholipid fatty acid analysis of adults with and without attention deficit/hyperactivity disorder.

Several psychiatric disorders, including juvenile Attention Deficit/Hyperactivity Disorder (ADHD), have been associated with abnormalities of certain long-chain PUFA (LCPUFA). Despite this reported association, the FA levels of patients with the adult form of ADHD have not previously been evaluated. In this study we measured the total blood phospholipid FA concentrations in 35 control subjects and 37 adults with ADHD symptoms to determine whether adults with ADHD symptoms would show abnormalities of FA relative to control subjects. In the serum phospholipids, adults with ADHD symptoms had significantly lower levels of total saturated, total polyunsaturated, and total omega-6 (n-6) FA, as well as the omega-3 (n-3) LCPUFA DHA (22:6n-3), and significantly higher levels of total monounsaturated FA and the n-3 LCPUFA docosapentaenoic acid (22:5n-3). In the erythrocyte membrane phospholipids, adults with ADHD symptoms had significantly lower levels of total PUFA, total n-3 FA, and DHA, and significantly higher levels of total saturated FA. Neither serum nor erythrocyte membrane phospholipid DHA was related to ADHD symptom severity (as assessed by the Amen questionnaire) in ADHD subjects. Although the exact cause of these variations is unknown, both environmental and genetic factors may be involved.